Fig. 2: SINEs inhibit the nuclear export of Cas9 mRNA in an XPO1-dependent manner. | Communications Biology

Fig. 2: SINEs inhibit the nuclear export of Cas9 mRNA in an XPO1-dependent manner.

From: KPT330 improves Cas9 precision genome- and base-editing by selectively regulating mRNA nuclear export

Fig. 2

a KPT330 reduces cytoplasmic Cas9 mRNA, as determined by the ratio of cytoplasmic Cas9 mRNA to total Cas9 mRNA using RT-qPCR. Significant difference is determined using Student’s t-test. b SINEs reduce cytoplasmic Cas9 mRNA in a dose-dependent manner, as determined by RT-qPCR. mRNA expression is normalized to drug-free group. a, b β-actin is used as an internal control. c Screening siRNA screen for XPO1 knockdown. d Evaluation of the efficiencies of siRNA knockdown of XPO1 mRNA expression in the absence and presence of 0.5 μM KPT330, as determined by RT-qPCR. e The effects of XPO1 knockdown on Cas9 mRNA transport in the absence and presence of 0.5 μM KPT330. d, e Significant difference between sham and XPO1 knockdown is determined using Student’s t-test. f RIP experiment showing HuR and LRPPRC binding with Cas9 mRNA. g The effects of KPT330 treatment on the expression of XPO1, HuR and LRPPRC proteins. h Cartoon illustrating SINE-mediated modulation of Cas9 mRNA transport. The above data are shown as mean ± SD (n = 2 or 3 biologically independent samples).

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